Effect of cross-trial nonstationarity on joint-spike events.

Common to most correlation analysis techniques for neuronal spiking activity are assumptions of stationarity with respect to various parameters. However, experimental data may fail to be compatible with these assumptions. This failure can lead to falsely assigned significant outcomes. Here we study the effect of nonstationarity of spike rate across trials in a model-based approach. Using a two-rate-state model, where rates are drawn independently for trials and neurons, we show in detail that nonstationarity across trials induces apparent covariation of spike rates identified as the generator of false positives. This finding has specific implications for the "shuffle predictor." Within the framework developed for our model, covariation of spike rates and the mechanism by which the shuffle predictor leads to wrong interpretation of the data can be discussed. Corrections for the influence of nonstationarity across trials by improvements of the predictor are presented.

Spike synchronization and firing rate in a population of motor cortical neurons in relation to movement direction and reaction time.

We studied the dynamics of precise spike synchronization and rate modulation in a population of neurons recorded in monkey motor cortex during performance of a delayed multidirectional pointing task and determined their relation to behavior. We showed that at the population level neurons coherently synchronized their activity at various moments during the trial in relation to relevant task events. The comparison of the time course of the modulation of synchronous activity with that of the firing rate of the same neurons revealed a considerable difference. Indeed, when synchronous activity was highest, at the end of the preparatory period, firing rate was low, and, conversely, when the firing rate was highest, at movement onset, synchronous activity was almost absent. There was a clear tendency for synchrony to precede firing rate, suggesting that the coherent activation of cell assemblies may trigger the increase in firing rate in large groups of neurons, although it appeared that there was no simple parallel shifting in time of these two activity measures. Interestingly, there was a systematic relationship between the amount of significant synchronous activity within the population of neurons and movement direction at the end of the preparatory period. Furthermore, about 400 ms later, at movement onset, the mean firing rate of the same population was also significantly tuned to movement direction, having roughly the same preferred direction as synchronous activity. Finally, reaction time measurements revealed a directional preference of the monkey with, once again, the same preferred direction as synchronous activity and firing rate. These results lead us to speculate that synchronous activity and firing rate are cooperative neuronal processes and that the directional matching of our three measures--firing rate, synchronicity, and reaction times--might be an effect of behaviorally induced network cooperativity acquired during learning.

Host defense against Pseudomonas aeruginosa requires ceramide-rich membrane rafts.

Pseudomonas aeruginosa infection is a serious complication in patients with cystic fibrosis and in immunocompromised individuals. Here we show that P. aeruginosa infection triggers activation of the acid sphingomyelinase and the release of ceramide in sphingolipid-rich rafts. Ceramide reorganizes these rafts into larger signaling platforms that are required to internalize P. aeruginosa, induce apoptosis and regulate the cytokine response in infected cells. Failure to generate ceramide-enriched membrane platforms in infected cells results in an unabated inflammatory response, massive release of interleukin (IL)-1 and septic death of mice. Our findings show that ceramide-enriched membrane platforms are central to the host defense against this potentially lethal pathogen.

Alternative medicine in allergies - prevalence, patterns of use, and costs.

BACKGROUND: There is evidence that the use of alternative medicine (AM) for allergies has increased. However, little is known from population-based studies about what determines its use. The objective of this study was to evaluate the patterns of use of AM for allergies. METHODS: A population-based nested case-control study was conducted in 2000-01 using computer-assisted telephone interviews. Three hundred and fifty-one adults participated (median age 46 years) with allergies including hay fever, asthma, atopic eczema, and food hypersensitivity. Information was obtained on demographics, prevalence, motivation, information, type of AM, provider, costs, willingness to pay, and subjective assessment of AM. RESULTS: 26.5% of participants used AM because of their allergies. Compared to nonusers, this group of users was significantly younger (median age 43 vs 47; p=0.004) and better educated (school education > 8 year vs

CD95 signaling via ceramide-rich membrane rafts.

Clustering seems to be employed by many receptors for transmembrane signaling. Here, we show that acid sphingomyelinase (ASM)-released ceramide is essential for clustering of CD95. In vitro and in vivo, extracellularly orientated ceramide, released upon CD95-triggered translocation of ASM to the plasma membrane outer surface, enabled clustering of CD95 in sphingolipid-rich membrane rafts and apoptosis induction. Whereas ASM deficiency, destruction of rafts, or neutralization of surface ceramide prevented CD95 clustering and apoptosis, natural ceramide only rescued ASM-deficient cells. The data suggest CD95-mediated clustering by ceramide is prerequisite for signaling and death.

Invasion of human epithelial cells by Pseudomonas aeruginosa involves src-like tyrosine kinases p60Src and p59Fyn.

Pseudomonas aeruginosa plays a major role in respiratory tract infections or sepsis in patients with cystic fibrosis or upon suppression of the immune system. Several P. aeruginosa strains have been shown to be internalized by human epithelial cells; however, the molecular mechanisms of the invasion process are poorly characterized. Here, we show that the internalization of P. aeruginosa into human epithelial cells results in and requires activation of the Src-like tyrosine kinases p59Fyn and p60Src and the consequent tyrosine phosphorylation of several eukaryotic proteins. The significance of Src-like tyrosine kinase activation is shown by an almost complete blockade of P. aeruginosa internalization, but not adhesion, upon inhibition of Src-like tyrosine kinases. Likewise, inhibition of P. aeruginosa binding to CFTR, which has been shown to block P. aeruginosa internalization, prevents Src and Fyn activation, supporting a pivotal role of Src-like tyrosine kinases for invasion by P. aeruginosa.

CD95/CD95 ligand interactions on epithelial cells in host defense to Pseudomonas aeruginosa.

Pseudomonas aeruginosa causes severe infections, particularly of the lung, that are life threatening. Here, we show that P. aeruginosa infection induces apoptosis of lung epithelial cells by activation of the endogenous CD95/CD95 ligand system. Deficiency of CD95 or CD95 ligand on epithelial cells prevented apoptosis of lung epithelial cells in vivo as well as in vitro. The importance of CD95/CD95 ligand-mediated lung epithelial cell apoptosis was demonstrated by the rapid development of sepsis in CD95- or CD95 ligand-deficient mice, but not in normal mice, after P. aeruginosa infection.

CD95-mediated apoptosis in vivo involves acid sphingomyelinase.

Acid sphingomyelinase (ASM) is reported to have an essential function in stress-induced apoptosis although the physiological function of ASM in receptor-triggered apoptosis is unknown. Here, we delineate a pivotal role for ASM in CD95-triggered apoptosis of peripheral lymphocytes or hepatocytes in vivo. We employed intravenous injection of anti-CD4 antibodies or phytohemagglutinin that was previously shown to result in apoptosis of peripheral blood lymphocytes or hepatocytes via the endogenous CD95/CD95 ligand system. Our results demonstrate a high susceptibility in normal mice whereas ASM knock-out mice fail to immunodeplete T cells or develop autoimmune-like hepatitis. Likewise, ASM-deficient mice or hepatocytes and splenocytes ex vivo manifest resistance to anti-CD95 treatment. These results provide in vivo evidence for an important physiological function of ASM in CD95-induced apoptosis.

Dynamical changes and temporal precision of synchronized spiking activity in monkey motor cortex during movement preparation.

Movement preparation is considered to be based on central processes which are responsible for improving motor performance. For instance, it has been shown that motor cortical neurones change their activity selectively in relation to prior information about movement parameters. However, it is not clear how groups of neurones dynamically organize their activity to cope with computational demands. The aim of the study was to compare the firing rate of multiple simultaneously recorded neurones with the interaction between them by describing not only the frequency of occurrence of epochs of significant synchronization, but also its modulation in time and its changes in temporal precision during an instructed delay. Multiple single-neurone activity was thus recorded in monkey motor cortex during the performance of two different delayed multi-directional pointing tasks. In order to detect conspicuous spike coincidences in simultaneously recorded spike trains by tolerating temporal jitter ranging from 0 to 20 ms and to calculate their statistical significance, a modified method of the 'Unitary Events' analysis was used. Two main results were obtained. First, simultaneously recorded neurones synchronize their spiking activity in a highly dynamic way. Synchronization becomes significant only during short periods (about 100 to 200 ms). Several such periods occurred during a behavioural trial more or less regularly. Second, in many pairs of neurones, the temporal precision of synchronous activity was highest at the end of the preparatory period. As a matter of fact, at the beginning of this period, after the presentation of the preparatory signal, neurones significantly synchronize their spiking activity, but with low temporal precision. As time advances, significant synchronization becomes more precise. Data indicate that not only the discharge rate is involved in preparatory processes, but also temporal aspects of neuronal activity as expressed in the precise synchronization of individual action potentials.

Precise spike synchronization in monkey motor cortex involved in preparation for movement.

It is commonly accepted that perceptually and behaviorally relevant events are reflected in changes of activity in largely distributed neuronal populations. However, it is much less clear how these populations organize dynamically to cope with momentary computational demands. In order to decipher the dynamic organization of cortical ensembles, the activities of up to seven neurons of the primary motor cortex were recorded simultaneously. A monkey was trained to perform a pointing task in six directions. During each trial, two signals were presented consecutively. The first signal provided prior information about the movement direction, whereas the second called for the execution of that movement. Dynamic interactions between the activity of simultaneously recorded neurons were studied by analyzing individual epochs of synchronized firing ("unitary events"). Unitary events were defined as synchronizations which occur significantly more often than expected by chance on the basis of the neurons' firing rates. The aim of the study was to describe the relationships between synchronization dynamics and changes in activity of the same neurons during the preparation and execution of voluntary movements. The data show that even neurons which were classified, on the basis of the change in their firing rate, to be functionally involved in different processes (e.g., preparation or execution related, different directional tuning) synchronized their spiking activity significantly. These findings indicate that the synchronization of individual action potentials and the modulation of the firing rate may serve different and complementary functions underlying the cortical organization of cognitive motor processes.

The distribution of neuronal population activation (DPA) as a tool to study interaction and integration in cortical representations.

In many cortical areas, simple stimuli or task conditions activate large populations of neurons. We hypothesize that such populations support processes of interaction within parametric representations and integration of multiple sources of input and we propose to study these processes using distributions of population activation (DPAs) as a tool. Such distributions can be viewed as neuronal representations of continuous stimulus or task parameters. They are built from basis functions contributed by each neuron. These functions may be explicitly chosen based on tuning curves or receptive field profiles. Or they may be determined by minimizing the distance between chosen target distributions and the constructed DPAs. In both cases, construction of the DPA is based on a set of reference conditions in which the stimulus or task parameters are sampled experimentally. In a second step, basis functions are kept fixed, and the DPAs are used to explore time dependent processing, interaction and integration of information. For instance, stimuli which simultaneously specify multiple parameter values can be used to study interactions within the parametric representation. We review an experiment, in which the representation of retinal position is probed in this way, revealing fast excitatory interactions among neurons representing similar retinal positions and slower inhibitory interactions among neurons representing dissimilar retinal positions. Similarly, DPAs can be used to analyze different sources of input that are fused within a parametric representation. We review an experiment in which the representation of the direction of goal-directed arm movements in motor and premotor cortex is studied when prior and current information about upcoming movement tasks are integrated.

Detecting unitary events without discretization of time.

In earlier studies we developed the 'Unitary Events' analysis (Grün S. Unitary Joint-Events in Multiple-Neuron Spiking Activity: Detection, Significance and Interpretation. Reihe Physik, Band 60. Thun, Frankfurt/Main: Verlag Harri Deutsch, 1996.) to detect the presence of conspicuous spike coincidences in multiple single unit recordings and to evaluate their statistical significance. The method enabled us to study the relation between spike synchronization and behavioral events (Riehle A, Grün S, Diesmann M, Aertsen A. Spike synchronization and rate modulation differentially involved in motor cortical function. Science 1997;278:1950-1953.). There is recent experimental evidence that the timing accuracy of coincident spiking events, which might be relevant for higher brain function, may be in the range of 1-5 ms. To detect coincidences on that time scale, we sectioned the observation interval into short disjunct time slices ('bins'). Unitary Events analysis of this discretized process demonstrated that coincident events can indeed be reliably detected. However, the method looses sensitivity for higher temporal jitter of the events constituting the coincidences (Grün S. Unitary Joint-Events in Multiple-Neuron Spiking Activity: Detection, Significance and Interpretation. Reihe Physik, Band 60. Thun, Frankfurt/Main: Verlag Harri Deutsch, 1996.). Here we present a new approach, the 'multiple shift' method (MS), which overcomes the need for binning and treats the data in their (original) high time resolution (typically 1 ms, or better). Technically, coincidences are detected by shifting the spike trains against each other over the range of allowed coincidence width and integrating the number of exact coincidences (on the time resolution of the data) over all shifts. We found that the new method enhances the sensitivity for coincidences with temporal jitter. Both methods are outlined and compared on the basis of their analytical description and their application on simulated data. The performance on experimental data is illustrated.

Prior information preshapes the population representation of movement direction in motor cortex.

Single neuron activity was recorded in monkey motor cortex during the execution of pointing movements in six directions. The amount of prior information was manipulated by varying the range of precued directions. A distribution of neural population activation was constructed in the space of movement directions. This population representation of movement direction was preshaped by the precue. Peak location and width reflected the precued range of movement directions. From this preshaped form, the population representation evolved continuously in time and gradually in parameter space toward a more sharply peaked distribution centered on the parameter value specified by the response signal. A theoretical model of motor programming generated a similar temporal evolution of an activation field representing movement direction.

Analyzing Neuronal Processing Locus in Stimulus-Response Association Tasks

If a neuron is being recorded while a trained animal performs a 2x2 stimulus-response association task, how can we decide whether it is related more to the encoding and analysis of the sensory stimulus, to the preparation and execution of the motor response, or to the animal's decision that associates the two? The difficulty arises because, within a single task, stimulus and response are intrinsically confounded per task instruction; it is only through proper analysis of errors in performance (behavioral noise) and variance in recorded neural activity (neuronal noise) that one can identify the sensorimotor significance of such activity. A quantitative technique is proposed here, based on the framework of signal detection theory, to determine the sensorimotor "locus" of a neural process when recorded simultaneously with the animal's performance on a trial-by-trial basis. The premise is that a pure sensory process should be influenced only by the nature of the sensory stimulus regardless of the nature of the behavioral response, and vice versa for a pure motor process. From the recorded neural activity, we calculate the prediction or discriminability (by an ideal operator) for the stimulus categories and for the response categories. These discriminability values are then compared with each other to infer whether the neural process is more related to stimulus or to response. An index is derived that quantitatively specifies the processing locus of a given neural process along the sensorimotor continuum, with pure sensory and pure motor processes at the two extremes. In between lies the locus of decision-related processes whose activities allow equal (but not chance) prediction for stimulus and response categories. The technique is applied to single-unit activities recorded in monkey primary motor cortex (MI) while the monkey performed a simple go/nogo task involving visual stimulus and hand/wrist movement. We find that sensorimotor indices of MI neurons are widely distributed, with a preponderance of motor-related units (that better predict go/nogo response than go/nogo stimulus) but also sensory-related ones (with predictabilities reversed). Copyright 1997 Academic Press

Dynamics of single neuron activity in monkey primary motor cortex related to sensorimotor transformation.

We investigated the dynamics of neuronal activity related to sensorimotor transformation during single experimental trials of a given stimulus-response (S-R) association task. A monkey was trained to perform wrist extension/flexion movements in the horizontal plane to align a pointer with a visual target while single unit activity in the primary motor cortex (MI) was being recorded. The stimulus was a colored light-emitting diode (LED) presented to either the left or right of a central reference point. The monkey had to point directly at the target ("compatible" S-R mapping) or point to the opposite side of the target position ("incompatible" S-R mapping), with the mapping rule specified by the color of the LED. Single neuron activities on the four correct trials (left/right stimulus x compatible/incompatible S-R mapping) were compared to determine whether such activities were more related to stimulus encoding and representation, to response preparation and execution, or to the "decision" processes translating the stimulus representation into a response representation. A novel mathematical technique, called LOCUS ANALYSIS, has been developed to quantitatively analyze and visualize the contribution of neuronal activity toward the sensory, motor, or sensorimotor (i.e., decisional) aspects of the task. Our data show that as a trial evolves, neuronal activity in MI, at a population level, is first correlated with the representation of the specific stimulus (the side of LED), then with the representation of the S-R mapping rule (the color of LED) as well as trial-specific S-R association (the conjunction of stimulus side and stimulus color), and finally with the representation of the behavioral response (extension or flexion wrist movement). Immediately after the issuance of the movement command, the populational activity in MI remains correlated with the trial-specific stimulus-response conjunctions, i.e., the context of the motor decision that the monkey has just made. Cells recorded successively in a single penetration tend to resemble each other in their pattern of firing on the four correct trials, suggesting a modular organization of neurons based on their functional role in the processing of the S-R association task. Our results indicate that MI belongs to a distributed network such that its neuronal activity reflects the underlying network dynamics that translate a stimulus representation into a response representation via the activation and application of appropriate S-R mapping rule.

Spike synchronization and rate modulation differentially involved in motor cortical function.

It is now commonly accepted that planning and execution of movements are based on distributed processing by neuronal populations in motor cortical areas. It is less clear, though, how these populations organize dynamically to cope with the momentary computational demands. Simultaneously recorded activities of neurons in the primary motor cortex of monkeys during performance of a delayed-pointing task exhibited context-dependent, rapid changes in the patterns of coincident action potentials. Accurate spike synchronization occurred in relation to external events (stimuli, movements) and was commonly accompanied by discharge rate modulations but without precise time locking of the spikes to these external events. Spike synchronization also occurred in relation to purely internal events (stimulus expectancy), where firing rate modulations were distinctly absent. These findings indicate that internally generated synchronization of individual spike discharges may subserve the cortical organization of cognitive motor processes.

Neuronal correlates of sensorimotor association in stimulus-response compatibility.

Neuronal mechanisms underlying stimulus-response (S-R) associations in S-R compatibility tasks were identified in 2 experiments with monkeys. Visual stimuli were presented on the left and right calling for left-right movements under congruent and incongruent S-R mapping instructions. High- and low-pitched tones calling for left-right movements were presented to the left and right ear, and the stimulus side was irrelevant. Single neurons sensitive to the S-R mapping rule were found in the primary motor cortex. The large overlap between the neuronal populations sensitive to the stimulus side, the S-R mapping rule, and the response side, respectively, is consistent with the idea that sensory-to-motor transformation is a continuous rather than a discrete process. Results partly support the hypothesis that the increase in reaction time with incongruent mapping is caused by the automatic activation of the congruent, but erroneous, response.

Neural correlates of partial transmission of sensorimotor information in the cerebral cortex.

Using single neuron recordings in monkey primary motor (MI) cortex, two series of experiments were conducted in order to know whether response preparation can begin before perceptual processing finishes, thus providing evidence for a temporal overlap of perceptual and motor processes. In Experiment 1, a "left/right, Go/No-Go" reaction time (RT) task was used. One monkey was trained to perform wrist flexion/extension movements to align a pointer with visual targets. The visual display was organized to provide a two-dimensional stimulus: side (an easy discrimination between left and right targets) which determined movement direction, and distance (a difficult discrimination between distal and proximal targets) which determined whether or not the movement was to be made. Changes in neuronal activity, when they were time-locked to the stimulus, were almost similar in the Go and No-Go trials, and when they were time-locked to movement onset, were markedly reduced in No-Go as compared to Go trials. In Experiment 2, a stimulus-response compatibility (SRC) task was used. Two monkeys were trained to align a pointer with visual targets, on either left or right. In the spatially "compatible" trials, they had to point at the stimulus position, whereas in the "incompatible" trials, they had to point at the target located in the opposite side. For 12.5% of neurons, changes in activity associated with incompatible trials looked like changes in activity associated with movements performed in the opposite direction during compatible trials, thus suggesting the hypothesis of an automatic activation of the congruent, but incorrect response. Results of both experiments provide evidence for a partial transmission of information from visual to motor cortical areas: that is, in the No-Go trials of the first task, information about movement direction, before the decision to perform or not this movement was made, and, in the incompatible trials of the SRC task, information about the congruent, but incorrect response, before the incongruent, but correct response was programmed.

Neuronal correlates of the specification of movement direction and force in four cortical areas of the monkey.

Single-neuron activity was recorded in several areas of the cerebral cortex when monkeys performed a movement-precueing reaction time task. In such a task, information provided by a first signal ('preparatory signal', PS) refers to what has to be done in response to a second signal ('response signal', RS). Two monkeys were trained to rotate a handle by performing wrist flexion/extension movements while two levels of frictional resistance were applied to the manipulandum. The PS provided complete, partial or no prior information about movement direction (flexion or extension) and/or the level of the frictional force (weak or strong). Since providing partial prior information about either movement parameter shortened reaction time (RT)--RT being shorter when movement direction than movement force was precued--, as compared to the condition in which no prior information was provided, the analysis of changes in neuronal activity during the preparatory period (PP), i.e., the instructed delay between PS and RS, makes the study of the neuronal mechanism underlying the specification of movement parameters possible. The activity of 411 neurons of the primary motor (MI), premotor (PM), somatosensory (SI) and parietal (PA) cortex was recorded during task performance. Many more neurons changed selectively their activity in relation to movement direction than in relation to movement force, not only during PP, but also during RT and movement time (MT). The number of purely direction-related neurons increased, whereas the number of purely force-related neurons decreased from SI to PA, then to MI and finally to PM. During PP, selective activity changes were related only to one movement parameter, whereas during RT and MT, a large population of neurons changed its activity in relation to both movement direction and force, especially in MI. These data provide further evidence for the clustering of distinct neuronal populations responsible for programming movement direction and force.

Neuronal coding of stimulus-response association rules in the motor cortex.

Two monkeys were trained to perform wrist movements to align a pointer with visual targets. In the spatially 'compatible' condition, monkeys had to point at the target position (left/right), whereas in the 'incompatible' condition, they had to point at the position opposite to the target. A large proportion of neurones recorded in the primary motor cortex showed changes in activity according to either the side of the target or the side of the movement. However, more than 40% of neurones changed their activity as a function of the stimulus-response mapping rule. Some of these neurones, being sensitive only to the stimulus-response compatibility effect, must therefore be viewed as specifically involved in the neural mechanisms that control the association process between sensory inputs and motor outputs.